Microdomain Ca2+ dynamics in mammalian muscle following prolonged high pressure treatments

Abstract

High pressure (HP) applications are an important thermodynamic tool to influence cellular processes. Especially processes that undergo large volume changes, e.g. opening or closing of ion channels, are in particular susceptible to HP treatments. Such volume changes are extremely difficult to assess for intracellular ion channels, like ryanodine receptors (RyR) residing in the membrane of organelles. In skeletal muscle, RyR act as Ca2+ release channels. We previously showed that plasmalemmal Na+ and Ca2+ ion channels were irreversibly altered after prolonged 20 MPa treatments. Here, changes in microdomain Ca2+ levels due to elementary Ca2+ release events (ECRE) were monitored using confocal fluorescence microscopy. We studied ECRE in mammalian skeletal muscle following 3 h HP treatments up to 30 MPa to clarify whether RyR induced intracellular microdomain Ca2+ dynamics was more susceptible to HP treatment compared to surface membrane ion currents. ECRE frequencies exponentially declined with pressure. ECRE amplitudes and rise times (RT) were quite robust towards HP treatments. In contrast, spatial and temporal ECRE extension showed a tendency towards larger values up to 20 MPa but declined for higher pressures. Activation volumes for pressure-induced persistent ECRE alterations were zero for RT but showed a bimodal behavior for event duration. It seems that although ECRE frequencies are markedly reduced, ECRE morphology is less affected by HP. In particular, RyR opening time is practically unaltered and the observed morphological ECRE changes might reflect alterations in local Ca2+ buffers and Ca2+ concentration profiles rather than involvement of RyR in mammalian skeletal muscle.